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DNA synthesis and induced mutations in the presence of 5-bromouracil II. Induction of mutations

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Summary

An investigation has been carried out on the heritable changes caused by the incorporation of 5-bromouracil (BU) into the DNA of a number of amino acid-requiring derivatives of a thymineless mutant ofEscherichia coli 15. Results revealed that the number of mutations induced by the analogue was linearly related to the amount of BU incorporated into DNA. DNA replication was necessary in the presence of BU for the induction of mutations, but concomitant RNA and protein synthesis were not necessary. Induced revertants were found to arise in either one of two distinct patterns: 1. in a single burst one replication after incorporation of BU, with no further increase upon subsequent DNA replication, and 2. continually with each additional DNA replication. Based on theFreese theory, these patterns allowed tentative identification of the base pairs involved. Further, the number of DNA replications necessary for the expression of BU induced reversions was determined. It was found that a transition from A:T to G:C required two DNA replications in addition to incorporation of BU, while the transition C:G to A:T required only one. Results obtained by the use of BU were compared with results obtained using AP. It was found that the pattern of mutant induction and the number of DNA replications necessary for the expression of each mutant were the same with AP as with BU. This implied that AP preferentially replaces adenine. As the number of DNA replications necessary for the expression of BU-and AP-induced reversions was the same for each mutant tested, it was concluded that both strands of the DNA molecule were necessary to carry the genetic message. An attempt was made to synchronize the replication of DNA and, thereby to synchronize the appearance of mutations during one replication cycle. Revertants, however, occurred uniformly throughout the cycle. The results were further discussed in relation to existing models of DNA replication.

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With 10 Figures in the Text

Supported in part by grants from the American Cancer Society, the U.S. Public Health Service and the National Science Foundation administered by ProfessorF. J. Ryan.

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Strelzoff, E. DNA synthesis and induced mutations in the presence of 5-bromouracil II. Induction of mutations. Zeitschrift für Vererbungslehre 93, 301–318 (1962). https://doi.org/10.1007/BF00889172

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